Lipolytic production from solid-state fermentation of the filamentous fungus Penicillium polonicum and its applicability as biocatalyst in the synthesis of ethyl oleate.

Lipases represent versatile biocatalysts extensively employed in transesterification reactions for ester production. Ethyl oleate holds significance in biodiesel production, serving as a sustainable alternative to petroleum-derived diesel. In this study, our goal was to prospect lipase and assess its efficacy as a biocatalyst for ethyl oleate synthesis. For quantitative analysis, a base medium supplemented with Rhodamine B, olive oil, and Tween 80 was used. Solid-state fermentation utilized crambe seeds of varying particle sizes and humidity levels as substrates. In the synthesis of ethyl oleate, molar ratios of 1:3, 1:6, and 1:9, along with a total enzymatic activity of 60 U in n-heptane, were utilized at temperatures of 30 °C, 37 °C, and 44 °C. Reactions were conducted in a shaker at 200 rpm for 60 min. As a result, we first identified Penicillium polonicum and employed the method of solid-state fermentation using crambe seeds as a substrate to produce lipase. Our findings revealed heightened lipolytic activity (22.5 Ug-1) after 96 h of fermentation using crambe cake as the substrate. Optimal results were achieved with crambe seeds at a granulometry of 0.6 mm and a fermentation medium humidity of 60%. Additionally, electron microscopy suggested the immobilization of lipase in the substrate, enabling enzyme reuse for up to 4 cycles with 100% enzymatic activity. Subsequently, we conducted applicability tests of biocatalysts for ethyl oleate synthesis, optimizing parameters such as the acid/alcohol molar ratio, temperature, and reaction time. We attained 100% conversion within 30 min at 37 °C, and our results indicated that the molar ratio proportion did not significantly influence the outcome. These findings provide a methodological alternative for the utilization of biocatalysts in ethyl oleate synthesis.

Functional responses of stream fish communities to rural and urban land uses

We tested the effects of ruralization and urbanization on the functional diversity indices and the composition of functional traits of Neotropical stream fish communities. The study was carried out in 24 streams of the Pirapó, Piquiri, Paraná III and Iguassu river basins. Land use in the watershed was categorized as percentages of native vegetation, rural occupation and urban occupation. Statistical tests revealed negative bivariate correlations between functional dispersion and the proportion of native vegetation in the watershed. The results indicate that a higher percentage of rural or urban occupation is associated with increased functional dispersion. In the analyzes of trait composition, significant alterations were observed in response to urbanization while only the increase in herbivory responded to ruralization. As the area of native vegetation is reduced by urbanization, the trait composition changes, with reduced proportions of species with intolerance to hypoxia, migratory reproductive behavior, external fertilization, and subterminal mouth, and increased proportions of species with parental care, detritivory, internal fertilization, and an upper mouth. Therefore, fish species that have these specific characteristics are more likely to disappear from streams as urbanization progresses. In summary, urbanization was related to a greater change in the composition of functional traits than ruralization.(AU)

Nós testamos os efeitos da ruralização e da urbanização sobre os índices de diversidade funcional e da composição de traços funcionais em assembleias de peixes de riachos Neotropicais. Amostras foram feitas em 24 riachos distribuídos nas bacias dos rios Pirapó, Piquiri, Paraná III e Iguaçu. O uso do solo foi categorizado por meio das porcentagens de vegetação, ocupação rural e urbana. Testes estatísticos revelaram correlações negativas bivariadas entre a dispersão funcional e a proporção de vegetação. Os resultados indicaram que maior percentual de ocupação rural ou urbana está associado ao aumento da dispersão funcional. Nas análises de composição de traços foram observadas alterações significativas em resposta à urbanização, enquanto apenas o aumento de herbívoros respondeu à ruralização. À medida que a área de vegetação é reduzida, a composição de traços muda, com redução nas proporções de espécies com intolerância à hipóxia, comportamento reprodutivo migratório, fertilização externa e boca subterminal, e aumento da proporção daquelas com cuidado parental, detritivoria, fertilização interna e boca superior. Portanto, espécies que apresentam essas características têm maior probabilidade de desaparecer dos riachos à medida que a urbanização avança. Em resumo, a urbanização foi relacionada a maior alteração na composição de traços funcionais do que a ruralização.(AU)

Arms as areas for larval development of migratory fish species in a Neotropical reservoir and the influence of rainfall over abundances.

The aim of the present study was to verify the use of the arms of the Itaipu Reservoir as areas of initial development for migratory fish species and to assess the relationship between rainfall and the spawning of migratory fish. Accordingly, fish larvae were collected from five arms of the reservoir from 2009 to 2016 using 0.5 mm plankton nets. Density was standardized as the number of larvae per 10 m3 filtered water, and the captured larval and juvenile specimens were identified at the lowest-possible taxonomic level. The larvae were also classified according to the degree of development and notochord flexion stage: larval vitelline, pre-flexion, flexion and post-flexion. To evaluate the distribution of larval abundance and the developmental stage along the longitudinal gradients of the arms, the data were evaluated using a set of nested linear models, following the AIC and Bayesian information criteria. In addition, an analysis of covariance was performed to investigate the influence of rainfall on the larval abundance of migratory species. During sampling, several species of economic and conservation interest such as Salminus brasiliensis and Pseudoplatystoma corruscans were collected. The larvae of the migratory fish taxa were captured from all sampled arms, which indicate them as areas of initial development. Nevertheless, it was observed that larval density increases from fluvial towards lacustrine zones inside the arms. Also, the present study verified that species, even in lentic environments, respond positively to rainfall stimuli in a manner similar to that exhibited by conspecifics in lotic environments. Such results reinforce the necessity of the protection of arms aiming at the conservation of this main group of species impaired by the construction of dams.

Unified Multimetric Index for the Evaluation of the Biological Condition of Streams in Southern Brazil Based on Fish and Macroinvertebrate Assemblages.

We developed MMI models that combine responses of fish and benthic macroinvertebrates for the evaluation of the biotic integrity of streams. The MMI was developed using a dataset covering stream sampling sites in the South of Brazil. Reference streams were identified based on the physical and chemical conditions and riparian vegetation. Thirty-four metrics were calculated and evaluated for their range, redundancy, and responsiveness to the environmental perturbation. We applied a robust approach to select the most sensitive metrics and MMI models based on the complexity and ability of the index in distinguishing impacted and reference sites. The four best MMI models selected are composed of different combinations of the eight metrics: % fish herbivorous, fish evenness, fish abundance, % macroinvertebrate shredder; % macroinvertebrate predator; % macroinvertebrate tolerant, % macroinvertebrate swimmer, and % macroinvertebrate burrower. All of the MMI models selected presented good performance in distinguishing reference streams from those impacted by different forms of land use. This study is one of the few attempts to use more than one biological assemblage in a single-multimetric index. Accordingly, we believe that the unified MMI we developed could be a useful tool to assist in the conservation and management of water resources in Neotropical regions, specially, in the implementation of ecological integrity tools more cost-effectively.

Comparison of fish and macroinvertebrates as bioindicators of Neotropical streams.

The biomonitoring of aquatic ecosystems in developing countries faces several limitations, especially related to gathering resources. The present study aimed at comparing the responses of fish and benthic macroinvertebrates to environmental change, to identify which group best indicates the differences between reference and impacted streams in southern Brazil. We determined reference and impacted sites based on physical and chemical variables of the water. For the analysis and comparison of biological responses, we calculated 22 metrics and submitted them to a discriminant analysis. We selected from this analysis only six metrics, which showed that the two studied assemblages respond differently to environmental change. A larger number of metrics were selected for macroinvertebrates than for fish in the separate analysis. The metrics selected for macroinvertebrates in the pooled analysis (i.e., fish and macroinvertebrates together) were different from those selected in the separate analysis for macroinvertebrates alone. However, the metrics selected for fish in the pooled analysis were the same selected in the separate analysis for fish alone. The macroinvertebrate assemblage was more effective for distinguishing reference from impacted sites. We suggest the use of macroinvertebrates as bioindicators of Neotropical streams, especially in situations in which time and money are short.

Optimization of the sperm:oocyte ratio and sperm economy in the artificial reproduction of Rhamdia quelen using fructose as a sperm motility modulator.

This research was conducted to evaluate the effects of fructose as a modulator of sperm motility and its effects on the reduction in number of sperm cells in IVF using cryopreserved Rhamdia quelen semen. Sperm activation occurred in solutions containing fructose (0.0, 0.9, 1.8, 2.7, 3.6 and 4.5%). The sperm motility rate, velocity and duration of sperm motility were assessed by polynomial regression analysis and grouped by the principal component analysis (PCA). Then, the oocytes were mixed with semen at proportions of 1×10(4), 3×10(4), 5×10(4), 7×10(4) and 9×10(4) for the sperm:oocyte ratio and fertilization was induced by the activation of gametes with the fructose-containing solutions. The fertilization, hatching and larval normality rate were evaluated by response surface protocol and were further grouped by PCA. All sperm variables were affected by the activating solutions, and the most desirable theoretical results for the rate of sperm motility were obtained when using a solution containing 2.85% fructose. In the IVF and incubation assays, there was an interactive effect between the motile sperm:oocyte ratio and the fructose concentration on the rates of oocyte fertilization, hatching and on the clustered index for reproductive success. The results suggest the possibility of reducing the sperm cells on IVF by 17.77% when using a solution containing 2.28% fructose. In conclusion, the use of solutions containing fructose at concentrations that maximize sperm movement allow the reduction of the motile sperm:oocyte ratio, thus promoting sperm metabolic efficiencies and contributing to the feasibility of using cryopreserved semen at a large-scale in IVF.

Delirium during Weaning from Mechanical Ventilation.

Background. We compare the incidence of delirium before and after extubation and identify the risk factors and possible predictors for the occurrence of delirium in this group of patients. Methods. Patients weaned from mechanical ventilation (MV) and extubated were included. The assessment of delirium was conducted using the confusion assessment method for the ICU and completed twice per day until discharge from the intensive care unit. Results. Sixty-four patients were included in the study, 53.1% of whom presented with delirium. The risk factors of delirium were age (P = 0.01), SOFA score (P = 0.03), APACHE score (P = 0.01), and a neurological cause of admission (P = 0.01). The majority of the patients began with delirium before or on the day of extubation. Hypoactive delirium was the most common form. Conclusion. Acute (traumatic or medical) neurological injuries were important risk factors in the development of delirium. During the weaning process, delirium developed predominantly before or on the same day of extubation and was generally hypoactive (more difficult to detect). Therefore, while planning early prevention strategies, attention must be focused on neurological patients who are receiving MV and possibly even on patients who are still under sedation.

Influence of prone positioning on premature newborn infant stress assessed by means of salivary cortisol measurement: pilot study.

OBJECTIVE: This study sought to assess the influence of prone positioning on the stress of newborn premature infants through the measurement of the salivary cortisol concentration and the evaluation of physiological and behavioral responses before and after changes in body positioning. METHODS: Saliva samples were collected from newborn infants at two different times: the first (corresponding to the baseline) after a period of 40 minutes during which the infants were not subjected to any manipulation and were placed in the lateral or supine position, and the second 30 minutes after placement in the prone position. Variables including heart rate, respiratory rate, peripheral oxygen saturation, and the Brazelton sleep score were recorded before, during, and at the end of the period in the prone position. RESULTS: The sample comprised 16 newborn premature infants (56.3% male) with a gestational age between 26 and 36 weeks, postnatal age between 1 and 33 days, birth weight of 935 to 3,050g, and weight at the time of intervention of 870 to 2,890g. During the intervention, six participants breathed room air, while the remainder received oxygen therapy. The median salivary cortisol concentration was lower in the prone position compared to baseline (0.13 versus 0.20; p=0.003), as was the median Brazelton sleep score (p=0.02). The average respiratory rate was lower after the intervention (54.88±7.15 versus 60±7.59; p=0.0004). The remainder of the investigated variables did not exhibit significant variation. CONCLUSION: Prone positioning significantly reduced the salivary cortisol level, respiratory rate, and Brazelton sleep score, suggesting a correlation between prone positioning and reduction of stress in preterm infants.

Influência do posicionamento em prona sobre o estresse no recém-nascido prematuro avaliada pela dosagem de cortisol salivar: um estudo piloto / Influence of prone positioning on premature newborn infant stress assessed by means of salivary cortisol measurement: pilot study

Objetivo: Avaliar a influência da postura em prona sobre o estresse no recém-nascido prematuro por meio da dosagem do cortisol salivar e da avaliação das respostas fisiológicas e comportamentais, antes e após o posicionamento. Métodos: Foi realizada a coleta de saliva em cada recém-nascido em dois momentos: o primeiro (correspondente ao basal), sem manipulação prévia por 40 minutos, em decúbito lateral ou supino; e o segundo, 30 minutos após o posicionamento em prona. A frequência cardíaca e respiratória, saturação periférica de oxigênio e escala de sono de Brazelton foram registradas antes, durante e ao final do posicionamento em prona. Resultados: Participaram do estudo 16 recém-nascidos prematuros (56,3% masculino) com idade gestacional de 26 a 36 semanas, com 1 a 33 dias de vida, e peso variando de 935 a 3.050g ao nascimento e de 870 a 2.890g no dia da intervenção. Durante o posicionamento, seis recém-nascidos estavam em ar ambiente e os demais recebiam oxigênio suplementar. A mediana dos níveis de cortisol salivar foi menor durante o posicionamento em prona comparativamente ao basal (0,13 e 0,20; p=0,003), assim como a do escore de sono de Brazelton (p=0,02). A média da frequência respiratória foi menor após a intervenção (54,88±7,15 e 60±7,59; p=0,0004). As demais variáveis analisadas não apresentaram variação significativa. Conclusão: O posicionamento em prona diminuiu significativamente os níveis de cortisol salivar, da frequência respiratória e do escore de sono de Brazelton, sugerindo a correlação entre essa postura e a diminuição do estresse nesses recém-nascidos. .

Objective: This study sought to assess the influence of prone positioning on the stress of newborn premature infants through the measurement of the salivary cortisol concentration and the evaluation of physiological and behavioral responses before and after changes in body positioning. Methods: Saliva samples were collected from newborn infants at two different times: the first (corresponding to the baseline) after a period of 40 minutes during which the infants were not subjected to any manipulation and were placed in the lateral or supine position, and the second 30 minutes after placement in the prone position. Variables including heart rate, respiratory rate, peripheral oxygen saturation, and the Brazelton sleep score were recorded before, during, and at the end of the period in the prone position. Results: The sample comprised 16 newborn premature infants (56.3% male) with a gestational age between 26 and 36 weeks, postnatal age between 1 and 33 days, birth weight of 935 to 3,050g, and weight at the time of intervention of 870 to 2,890g. During the intervention, six participants breathed room air, while the remainder received oxygen therapy. The median salivary cortisol concentration was lower in the prone position compared to baseline (0.13 versus 0.20; p=0.003), as was the median Brazelton sleep score (p=0.02). The average respiratory rate was lower after the intervention (54.88±7.15 versus 60±7.59; p=0.0004). The remainder of the investigated variables did not exhibit significant variation. Conclusion: Prone positioning significantly reduced the salivary cortisol level, respiratory rate, and Brazelton sleep score, suggesting a correlation between prone positioning and reduction of stress in preterm infants. .

Morphological differentiation among migratory fish species from the Paraná River basin / Diferenciação morfológica entre espécies de peixes migradores da bacia do Rio Paraná

This study characterized the morphological differences among migratory fish species from the Paraná River Basin. A total of 177 adult fish of Leporinus elongatus, Leporinus macrocephalus, Prochilodus lineatus, Salminus brasiliensis, Pimelodus maculatus, Pinirampus pirinampu, Pseudoplatystoma corruscans, Pterodoras granulosus, and Rhaphiodon vulpinus were sampled in the Canal da Piracema, a fish passage system at Itaipu Dam, Paraná River. The migratory species were analyzed through 22 morphometric measurements and fineness ratio to identify morphological patterns related to swimming performance. Four species groups were indicated by the Cluster analysis: I) R. vulpinus; II) P. corruscans; III) L. macrocephalus, L. elongatus, P. lineatus, P. maculatus, and S. brasiliensis - measurements of head height and anal fin length formed this group; and IV) P. granulosus and P. pirinampu - measurements of maximum body width and caudal peduncle width contributed to form this group. The morphometric variables that most contributed to the distinction were head length and mouth width for groups I and II. The species R. vulpinus and P. granulosus differed from other species, showing fineness ratios of 9.4 and 3.7, respectively. Leporinus elongatus, L. macrocephalus, P. lineatus, P. maculatus, and S. brasiliensis showed significant differences from other species, with ratios ranging from 4.57 to 5.19, indicating that these species may be better swimmers. Long-distance migratory species using the Piracema Canal to ascend upstream areas differed morphologically, morphological characteristics such as a narrow caudal peduncle and maximum body length, besides to the values of the fineness ratio, were essential to differentiate the migratory species.

Este estudo caracterizou as diferenças morfológicas entre espécies de peixes migradores da bacia do rio Paraná. Um total de 117 peixes adultos das espécies Leporinus elongatus, Leporinus macrocephalus, Prochilodus lineatus, Salminus brasiliensis, Pimelodus maculatus, Pinirampus pirinampu, Pseudoplatystoma corruscans, Pterodoras granulosus e Rhaphiodon vulpinus foram capturados no Canal da Piracema, um sistema de transposição para peixes na barragem de Itaipu, rio Paraná. As espécies migradoras foram analisadas através de 22 medidas morfométricas e razão fineza para identificar padrões morfológicos relacionados à capacidade natatória. Quatro grupos de espécies foram evidenciados pela Análise de Cluster: I) R. vulpinus; II) P. corruscans; III) L. macrocephalus, L. elongatus, P. lineatus, P. maculatus e S. brasiliensis, medidas de altura de cabeça e comprimento da nadadeira anal formaram este grupo; e IV) P. granulosus e P. pirinampu - medidas de largura máxima do corpo e largura do pedúnculo caudal contribuíram para a formação deste grupo. As variáveis morfométricas que mais contribuíram para distinção dos grupos I e II foram comprimento da cabeça e largura da boca. As espécies R. vulpinus e P. granulosus diferiram das demais, apresentando razão de fineza de 9,4 e 3,7, respectivamente. Leporinus elongatus, L. macrocephalus, P. lineatus, P. maculatus, S. brasiliensis diferiram significativamente das demais espécies com valores de razão fineza que variaram entre 4,57 a 5,19, indicando que estas espécies podem apresentar melhor desempenho natatório. As espécies migradoras de longa distância que utilizam o Canal da Piracema para ascender áreas a montante diferiram morfologicamente, características morfológicas como pedúnculo caudal estreito e largura máxima do corpo, bem como os valores de razão de fineza foram determinantes na diferenciação das espécies aqui analisadas.

Fish larvae assemblages in two floodplain lakes with different degrees of connection to the Paraná River, Brazil

The objective of this study was to assess the abundance, attributes of assemblages, and spatial and temporal distributions of fish larvae and their relationships with some abiotic variables in two floodplain lakes with different degrees of connection to the Paraná River in Ilha Grande National Park, PR, Brazil. Four sampling sites were chosen, two in each floodplain lake. Night samples were taken with plankton nets during three spawning seasons (monthly, from October to March) from 2001 to 2005. The highest diversity and abundance were recorded at Saraiva Lake, with 25 taxa being identified. In Xambrê Lake, only sedentary species were captured, and the most abundant species were Plagioscion squamosissimus and Hypophthalmus edentatus. The greatest abundance of larvae was found in the second spawning season. In the Saraiva Lake, the most abundant species were Moenkhausia aff. intermedia, Hyphessobrycon sp., and Bryconamericus stramineus, but larvae of known migratory species were also documented. In this lake, the greatest abundance of larvae was found in the third spawning. Larvae abundance was influenced by water temperature and conductivity. The high diversity and abundance recorded in Saraiva Lake may be a result of its connectivity with the Paraná River, and the low diversity and abundance observed at Xambrê Lake are likely due to its isolation from the river. This work shows the importance of these lagoons for fish development, for both sedentary and migratory species. Both lagoons may be considered to have extreme ecological importance and they are also extremely susceptible to impacts, so any careless disturbance may cause irreversible damage.(AU)

Este trabalho teve por objetivo avaliar a abundância, os atributos da assembleia e a distribuição espacial e temporal de larvas de peixes, bem como a relação dessas variáveis com alguns fatores abióticos, em duas lagoas com diferentes graus de conexão com o rio Paraná, Parque Nacional de Ilha Grande, PR, Brasil. As amostragens foram realizadas em quatro locais, dois em cada lagoa. As coletas foram realizadas mensalmente à noite durante três períodos de desova (outubro a março) de 2001 a 2005, com o auxílio de redes de plâncton. A maior diversidade e abundância foram verificadas na lagoa Saraiva, sendo identificados 25 táxons. Na lagoa Xambrê somente foram capturadas espécies sedentárias e as mais abundantes foram Plagioscion squamosissimus e Hypophthalmus edentatus. Nessa lagoa as maiores ocorrências foram verificadas no período II. Na lagoa Saraiva as maiores capturas foram de Moenkhausia aff. intermedia, Hyphessobrycon sp. e Bryconamericus stramineus, entretanto, larvas de espécies migradoras também foram registradas. Nessa lagoa, as maiores ocorrências foram no período III. A abundância de larvas foi influenciada pela temperatura da água e pela condutividade elétrica. A elevada diversidade e abundância registradas na lagoa Saraiva podem ser atribuídas a sua conectividade com o rio Paraná, enquanto que a baixa diversidade e abundância observadas na lagoa Xambrê podem estar relacionadas ao seu isolamento. Os resultados obtidos com este estudo demonstram a importância das lagoas marginais para o desenvolvimento inicial dos peixes, tanto para espécies sedentárias como migradoras. Ambas as lagoas podem ser consideradas de extrema importância ecológica e altamente susceptíveis a impactos, onde qualquer interferência sem conhecimentos prévios pode causar danos irreversíveis.(AU)